The oligomerization of a family of four genetically clustered human gastrointestinal mucins.
Mucins are synthesized and secreted by many epithelia. They are complex glycoproteins that offer cytoprotection. In their functional configuration, mucins form oligomers by a biosynthetic process that is poorly understood. A family of four human gastrointestinal mucin genes (MUC2, MUC5AC, MUC5B, and MUC6) is clustered to chromosome 11p15.5. To study oligomerization of these related mucins, we performed metabolic labeling experiments with [35S]amino acids in LS174T cells, and isolated mucin precursors by specific immunoprecipitations that were analyzed on SDS-PAGE. Each of the precursors of MUC2, MUC5AC, MUC5B, and MUC6 formed a single species of disulfide-linked homo-oligomer within 1 h after pulse labeling. Based on apparent molecular masses, these oligomeric precursors were most likely dimers. Inhibition of vesicular RER-to-Golgi transport, with brefeldin A and CCCP, did not affect the dimerization of MUC2 precursors, localizing dimerization to the RER. O-Glycosylation of MUC2 followed dimerization. Inhibition of N-glycosylation by tunicamycin retarded, but did not inhibit, dimerization, indicating that N-glycans play a role in efficient dimerization of MUC2 precursors. Based on sequence homology, the ability of MUC2, MUC5AC, MUC5B and MUC6 to dimerize most likely resides in their C-terminal domains. Thus, the RER-localized dimerization of secretory mucins likely proceeds by similar mechanisms, which is an essential step in the formation of the human gastrointestinal mucus-gels.
van Klinken
B J
BJ
Pediatric Gastroenterology and Nutrition, Academic Medical Center, Rm 68-260, University of Amsterdam, Amsterdam, The Netherlands.
Einerhand
A W
AW
Büller
H A
HA
Dekker
J
J
D002460:Cell Line; D002880:Chromosomes, Human, Pair 11 / Q000235:genetics; D019281:Dimerization; D018870:Endoplasmic Reticulum, Rough / Q000737:chemistry; D005752:Gastric Mucins / Q000737:chemistry / Q000235:genetics; D005753:Gastric Mucosa / Q000737:chemistry; D006031:Glycosylation; D006801:Humans; D007413:Intestinal Mucosa / Q000737:chemistry; D055271:Mucin 5AC; D055262:Mucin-2; D055257:Mucin-5B; D055272:Mucin-6; D009077:Mucins / Q000737:chemistry* / Q000235:genetics*; D005810:Multigene Family; D011487:Protein Conformation; D011498:Protein Precursors / Q000737:chemistry / Q000235:genetics
D005752:Gastric Mucins; C524625:MUC2 protein, human; C524678:MUC5AC protein, human; C524578:MUC5B protein, human; C524734:MUC6 protein, human; D055271:Mucin 5AC; D055262:Mucin-2; D055257:Mucin-5B; D055272:Mucin-6; D009077:Mucins; D011498:Protein Precursors